Excision of an integrated provirus by the action of FLP recombinase.
Retroviral vectors can be used to insert a single, intact copy of a transgene into a chromosome. If the duplication of the LTR (long-terminal repeat) that naturally occurs during reverse transcription of the retroviral genome is exploited to introduce two equally oriented FLP recognition target (FRT) sites, a substrate for FLP recombinase is created. A pulse of FLP recombinase activity can then be applied to excise the intervening sequences with the retention of a single LTR. This procedure is of potential use for manipulating an integration site after a period of expression enabling a variety of critical controls. We describe the properties of such a retroviral vector containing a dicistronic expression cassette with a reporter gene in the first and a positive/negative selection marker in the second cistron. This vector permits the selection and control of each step during the sequence of genomic manipulations enabled by site-specific recombination events.
Schübeler
D
D
Gesellschaft für Biotechnologische Forschung m.b.H., Generegulation und Differenzierung/Genetik von Eukaryonten, Mascheroder Weg, Germany.
Mielke
C
C
Bode
J
J
D016475:3T3 Cells; D000469:Alkaline Phosphatase / Q000235:genetics; D000818:Animals; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D018014:Gene Transfer Techniques; D005796:Genes; D017930:Genes, Reporter; D005822:Genetic Vectors; D051379:Mice; D016133:Polymerase Chain Reaction; D012194:RNA-Directed DNA Polymerase; D012091:Repetitive Sequences, Nucleic Acid; D012190:Retroviridae / Q000235:genetics*; D014771:Virion; D016662:Virus Integration
D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase; D012194:RNA-Directed DNA Polymerase; D000469:Alkaline Phosphatase